A promoter-trap vector pGBT14 for selecting promoters of fungus gene was constructed with E. coli-yeast shuttling plasmid pGBT9. Using this vector, a0. 5-2. 0kb chromosomal DNA library of Cepholosporium acremonium was constructed, and twenty four DNA fragments with promoter function in Saccharomyces oerevisiae Y153 were selected from this DNA library. And the promoter function of these DNA fragments was analyzed.

OBJECTIVETo observe and evaluate the phytoestrogenic effects and its mechanism of psoralen in estrogen receptor (ER) alpha and beta positive T47D and ishikawa cells.

METHODThe proliferation rate of T47D influenced by 1 x 10(-5) mol x L(-1) to 1 x 10(-9) mol x L(-1) psoralen and that of Ishikawa influenced by 1 x 10(-6) mol x L(-1) and 1 x 10(-7) mol x L(-1) psoralen were analyzed by MTT assay. PR mRNA expression in T47D was quantified by RT-PCR assay. Estrogen receptor antagonist ICI 182, 780 was employed as a tool. ER-alpha and ER-beta expression of T47D was measured by flow cytometry.

RESULTThe proliferation rates of T47D cells treated with 1 x 10(-5) mol x L(-1) to 1 x 10(-7) mol x L(-1) psoralen and ishikawa cells treated with 1 x 10(-6) mol x L(-1) to 1 x 10(-7) mol x L(-1) psoralen were increased significantly. The RT-PCR result showed that 1 x 10(-7) mol x L(-1) and 1 x 10(-6) mol x L(-1) psoralen could increase PR expression in T47D cells. The above effects could be blocked by ICI 182,780. Psoralen could also induce the augment of ER-alpha and ER-beta expression in T47D cells significantly.

CONCLUSIONPsoralen has phytoestrogenic effects. The effects are attained through ER pathway.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estradiol ; analogs & derivatives ; pharmacology ; Female ; Ficusin ; pharmacology ; Flow Cytometry ; Humans ; Receptors, Estrogen ; antagonists & inhibitors ; genetics ; Receptors, Progesterone ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Carnosol has been proved to have anti-breast cancer effect in previous research. But its ER subtype's specific regulation and mediation mechanisms remain unclear. The aim of this study is to observe the effect of carnosol on cell proliferation and its estrogen receptor α and β's specific regulation and mediation mechanisms with ER positive breast cancer T47D cell. With estrogen receptor α and β antagonists MPP and PHTPP as tools, the MTT cell proliferation assay was performed to observe the effect of carnosol on T47D cell proliferation. The changes in the T47D cell proliferation cycle were detected by flow cytometry. The effect of carnosol on ERα and ERβ expressions of T47D cells was measured by Western blot. The findings showed that 1 x 10(-5)-1 x 10(-7) mol x L(-1) carnosol could significantly inhibit the T47D cell proliferation, which could be enhanced by MPP or weakened by PHTPP. Meanwhile, 1 x 10(-5) mol x L(-1) or 1 x 10(-6) mol x L(-1) carnosol could significantly increase ERα and ERβ expressions of T47D cells, and remarkably increase ERα/ERβ ratio. The results showed that carnosol showed the inhibitory effect on the proliferation of ER positive breast cancer cells through target cell ER, especially ERβ pathway. In the meantime, carnosol could regulate expressions and proportions of target cell ER subtype ERα and ERβ.
Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Abietane ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Estrogen Receptor Modulators ; pharmacology ; Estrogen Receptor alpha ; antagonists & inhibitors ; metabolism ; Estrogen Receptor beta ; antagonists & inhibitors ; metabolism ; Female ; Flow Cytometry ; Humans ; Molecular Structure ; Pyrazoles ; pharmacology ; Pyrimidines ; pharmacology

OBJECTIVETo explore the phytoestrogenic effects of ten kinds of Chinese medicine including flos carthami, radix cyathulae, radix salviae miltiorrhizae, fructus ligustri lucidi, fructus lycii, radix clycyrrhizae, herba cistanches, herba epimedii, fructus psoraleae and semen cuscutae.

METHOD240 female Kunming mice weighting 9 - 12 g were randomly divided into two main groups A and B. A group was divided into 12 small groups: 1 solvent control group, 1 diethylstilbestrol control group and 10 Chinese medicine groups. B group was also divided into 12 small groups: 1 solvent control group, 1 diethylstilbestrol control group and 10 Chinese medicine antagonistic groups. Mice in ten antagonistic groups were administered both Chinese medicine and diethylstilbestrol everyday. After administered(op) for 4 days, blood was collected and serum was separated. The effect of the pharmacological serum on proliferation rate of MCF-7 (ER+) was analyzed by MTT-assay.

RESULTIn A group, proliferation rates of MCF-7 cells treated with serum from eight Chinese medicine groups including flos carthami, radix cyathulae, radix salviae miltiorrhizae, fructus lycii, herba cistanches, herba epimedii, fructus psoraleae and semen cuscutae were coued markedly increase respectively. While serum from fructus ligustri lucidi group could markedly decrease the proliferation rate of MCF-7 cells. In B group, the increased proliferation rate of MCF-7 cells caused by diethylstilbestrol was significantly reduced in seven Chinese medicine antagonistic groups including flos carthami, radix cyathulae, radix salviae miltiorrhizae, radix clycyrrhizae, herba epimedii, fructus psoraleae and semen cuscutae. While the increased proliferation rate could be markedly enhanced in herba cistanches group.

CONCLUSIONSix kinds of Chinese medicine such as flos carthami, radix cyathulae, radix salviae miltiorrhizae, herba epimedii, fructus psoraleae and semen cuscutae show both estrogenic effects (when administered indepently) and antiestrogenic effects (when administered together with diethylstilbestrol). Such bidirectional effects depends on the internal estrogen level.

Animals ; Breast Neoplasms ; metabolism ; pathology ; Carthamus tinctorius ; chemistry ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Diethylstilbestrol ; pharmacology ; Drug Antagonism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Estrogens, Non-Steroidal ; pharmacology ; Female ; Humans ; Mice ; Phytoestrogens ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Random Allocation ; Receptors, Estrogen ; metabolism ; Salvia miltiorrhiza ; chemistry ; Serum

The coordination and unification of Yin and Yang are the basis of normal human life activities. Along with the age growth and aging of the body, women will suffer from menopausal syndrome during menopause. In addition to the significant changes in the genital system, there are also pathological manifestations in estrogen target points including bone, nerve and cardiovascular systems, due to the imbalance of Yin and Yang. Besides the insufficiency of estrogen, the main cause of menopausal syndrome is the changes in the response of target organs to estrogen. In other words, the biological effects mediated by estrogen receptor(ER) alpha and beta subtypes in target cells are often different or even opposite; the changes of expression level and ratio of ERα and ERβ are also important causes for the abnormal estrogenic effects in target organs and the imbalance of Yin and Yang of the body. Therefore, on one hand, the therapeutic mechanism of drugs is ER-mediated estrogenic effect. On the other hand, the drugs have a regulatory effect on ER subtype expression in target cells and Yin-Yang state in target organs and even organisms, so as to cause further changes in the response of target cells to estrogen or estrogenic components, and exert its therapeutic effects. This paper reviews the pharmacological mechanism of gynecological traditional Chinese medicine in harmonizing Yin and Yang in estrogen-positive target cells and the clinical efficacy in the following aspects, including estrogen and its mechanism, the estrogenic effect of ER in traditional Chinese medicine and the mechanism of ER subtype in balancing Yin and Yang and mediating and regulating the main target tissues in menopausal syndrome treatment.
Estrogen Receptor alpha ; Estrogen Receptor beta ; Estrogens ; Female ; Humans ; Medicine, Chinese Traditional ; Yin-Yang

OBJECTIVETo analyses the clinical application of thoracodorsal artery perforator flaps (TAP).

METHODSWe used free or pedicled TAP flaps in 7 patients from Aug 2006 to April 2007, The age ranged from 7 to 42 years old, the perforator arteries was detected and labeled with a hand held Doppler flowmeter, the size of flaps ranged from 6 cm x 9 cm - 12 cm x 16 cm, the flaps designed with perforator artery included, all the flaps are based on the first perforator artery.

RESULTSAll the flaps survived well, no complication occurred with lowest donor site morbidity.

CONCLUSIONSThe thoracodorsal artery flap with latissimus dorsal muscle saved is a thin and reliable flaps with robust of blood supply, the flap can reduce significantly donor site morbidity and is a good choice for reconstructive surgery.

Adolescent ; Adult ; Back ; blood supply ; Child ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; blood supply ; Thoracic Arteries ; transplantation ; Young Adult

OBJECTIVEThe causal agent for SARS is considered as a novel coronavirus that has never been described both in human and animals previously. The stability of SARS coronavirus in human specimens and in environments was studied.

METHODSUsing a SARS coronavirus strain CoV-P9, which was isolated from pharyngeal swab of a probable SARS case in Beijing, its stability in mimic human specimens and in mimic environment including surfaces of commonly used materials or in household conditions, as well as its resistance to temperature and UV irradiation were analyzed. A total of 10(6) TCID50 viruses were placed in each tested condition, and changes of the viral infectivity in samples after treatments were measured by evaluating cytopathic effect (CPE) in cell line Vero-E6 at 48 h after infection.

RESULTSThe results showed that SARS coronavirus in the testing condition could survive in serum, 1:20 diluted sputum and feces for at least 96 h, whereas it could remain alive in urine for at least 72 h with a low level of infectivity. The survival abilities on the surfaces of eight different materials and in water were quite comparable, revealing reduction of infectivity after 72 to 96 h exposure. Viruses stayed stable at 4 degrees C, at room temperature (20 degrees C) and at 37 degrees C for at least 2 h without remarkable change in the infectious ability in cells, but were converted to be non-infectious after 90-, 60- and 30-min exposure at 56 degrees C, at 67 degrees C and at 75 degrees C, respectively. Irradiation of UV for 60 min on the virus in culture medium resulted in the destruction of viral infectivity at an undetectable level.

CONCLUSIONThe survival ability of SARS coronavirus in human specimens and in environments seems to be relatively strong. Heating and UV irradiation can efficiently eliminate the viral infectivity.

Environment ; Hot Temperature ; Humans ; Pharynx ; virology ; SARS Virus ; isolation & purification ; pathogenicity ; Survival Analysis ; Ultraviolet Rays

The study aimed to illuminate the role of G protein coupled estrogen receptor( GPER) and its mediated PI3 K/AKT signaling pathway in cryptotanshinone( CPT) induced apoptosis of breast cancer SKBR-3 cells,which is GPER positive and ER negative.The apoptosis rate of SKBR-3 cells was tested by Annexin V-FITC/PI staining and apoptosis effector caspase-3 was determined by Western blot. The key proteins in PI3 K/AKT signaling pathway mediated by GPER were detected by Western blot and immunofluorescence technique. Meanwhile,the agonist G1 and antagonist G15 of GPER and antagonist LY294002 of PI3 K were employed in the test to further clarify the effect of GPER and PI3 K/AKT pathway. The results indicated that the apoptosis rate was increased from 4. 7% to46. 1% and 69. 0% after treatment with 0,5,10 μmol·L~(-1) CPT for 48 h( P<0. 01). The expression of PI3 K,AKT and p-AKT were inhibited( P<0. 05 or P<0. 01),while caspase-3 level increased obviously after treatment with CPT( P<0. 01). Importantly,inhibitory effect of PI3 K/AKT signaling pathway by CPT was further enhanced by G1 and attenuated by G15. LY294002 also induced a further inhibition of expression of AKT and p-AKT. The mean fluorescence intensity of AKT and p-AKT could be decreased by CPT. Furthermore,CPT could downregulate GPER expression in SKBR-3 cells( P<0. 01),which could be inhibited by G1 and enhanced by G15.In conclusion,CPT could induce the apoptosis of ER negative and GPER positive breast cancer SKBR-3 cells and the molecular mechanism is related to its regulatory effect of GPER and its mediated PI3 K/AKT signaling pathway.
Apoptosis ; Breast Neoplasms ; Drugs, Chinese Herbal ; Humans ; Proto-Oncogene Proteins c-akt ; Receptors, Estrogen ; Receptors, G-Protein-Coupled ; Signal Transduction

Hyaluronic acid is widely used in biomaterials, cosmetics, clinical medicine and other fields due to its good biocompatibility, degradability, hydrophilicity, tumor targeting, viscosity and other characteristics. Pharmacodynamic activities of natural small molecular products which derived from traditional Chinese medicine (TCM) are significant, but their low solubility and poor targeting limit the clinical application. Based on supramolecular properties of hyaluronic acid, in this review, numerous studies were reviewed on the improvement of solubility, bioavailability, targeting and suitable dosage forms of small molecular compounds in TCM by domestic and foreign scholars using hyaluronic acid as carrier. It provides new ideas and inspirations for exploring the potential application value of small molecule compounds in TCM and even for the research and development of new drugs.